Methods and compositions using integrin-based therapeutics

Abstract

The present invention is directed to modified integrin proteins and methods and compositions using integrin-based therapeutics. In one embodiment, the modified integrins demonstrate increased occurrence or duration of the E−H+ integrin protein conformation. In another embodiment, the compounds of the present invention stabilize E−H+ integrin protein conformation, increasing the occurrence or duration of the E−H+ integrin protein conformation. In another embodiment, the compounds of the present invention inhibit binding of a ligand of an integrin. In yet a further embodiment, the present compounds increase cis binding of the integrin or signaling based thereon. The present compounds decrease the occurrence or duration of trans binding of the integrin or signaling based thereon. The modified integrins and compounds described herein may be used in methods of treating immune modulated diseases or inflammatory diseases or conditions.

1 . A compound comprising:
 a. a stabilizer of E−H+ integrin protein confirmation;   b. a modified integrin demonstrating E−H+ structure; or   c. a polynucleotide comprising a nucleotide sequence encoding a modified integrin demonstrating E−H+ structure.   
     
     
         2 . The compound of  claim 1 , wherein the stabilizer is selected from an antibody that stabilizes the E−H+ integrin structure, a fusion protein, a protein, and a small molecule. 
     
     
         3 . The compound of any of  claim 1  or  2 , wherein the stabilizer is an antibody. 
     
     
         4 . The compound according to  claim 1 , wherein the integrin is selected from a β2 integrin, an α4β1 integrin, an α4β7 integrin, an αEβ7 integrin, an αV integrin, or an αIIbβ3 integrin. 
     
     
         5 . The compound according to  claim 4 , wherein the β2 integrin is selected from an αLβ2 integrin, αMβ2 integrin, αxβ2 integrin, or αdβ2 integrin. 
     
     
         6 . The compound of  claim 1 , wherein the compound has anti-inflammatory properties. 
     
     
         7 . The compound of  claim 1 , wherein the compound inhibits trans integrin binding. 
     
     
         8 . The compound of  claim 1 , wherein the compound agonizes cis integrin binding. 
     
     
         9 . A pharmaceutical composition comprising the compound according to  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         10 . A method of increasing the duration or occurrence of E−H+ integrin structure. 
     
     
         11 . A method of increasing the occurrence or duration of cis integrin binding and/or signaling comprising contacting a cell expressing an integrin with:
 a. a stabilizer of E−H+ integrin protein confirmation;   b. a modified integrin demonstrating E−H+ structure; or   c. a polynucleotide comprising a nucleotide sequence encoding a modified integrin demonstrating E−H+ structure.   
     
     
         12 . A method of treating an immune modulated disease and/or an inflammatory disease or condition disease comprising:
 administering an effective amount of the pharmaceutical composition according to any one of  claims 1  to  9  to a patient in need thereof.   
     
     
         13 . The method according to  claim 12 , wherein the immune modulated disease is selected from: multiple sclerosis, experimental autoimmune encephalomyelitis (both relapsing and remitting), rheumatoid arthritis, diabetes, eczema, psoriasis, the inflammatory bowel diseases, allergic disorders anaphylactic hypersensitivity, asthma, allergic rhinitis, atopic dermatitis, vernal conjunctivitis, eczema, urticarial, food allergies, allergic encephalomyelitis, multiple sclerosis, insulin-dependent diabetes mellitus, and autoimmune uveoretinitis, inflammatory bowel disease, Crohn's disease, regional enteritis, distal ileitis, granulomatous enteritis, regional ileitis, terminal ileitis, ulcerative colitis, autoimmune thyroid disease, hypertension, infectious diseases, allograft rejection (such as graft vs host disease), airway hyper reactivity, atherosclerosis, inflammatory liver disease, and cancer. 
     
     
         14 . The method according to  claim 13 , wherein the immune modulated disease is characterized by inflammation. 
     
     
         15 . The method according to  claim 12 , wherein the inflammatory disease or condition is selected from:
 general chronic or acute inflammation, inflammatory skin diseases, immune-related disorders, burn, immune deficiency, acquired immune deficiency syndrome (AIDS), myeloperoxidase deficiency, Wiskott-Aldrich syndrome, chronic kidney disease, chronic granulomatous disease, hyper-IgM syndromes, leukocyte adhesion deficiency, iron deficiency, Chediak-Higashi syndrome, severe combined immunodeficiency, diabetes, obesity, hypertension, HIV, wound-healing, remodeling, scarring, fibrosis, stem cell therapies, cachexia, encephalomyelitis, multiple schlerosis, psoriasis, lupus, rheumatoid arthritis, immune-related disorders, radiation injury, transplantation, cell transplantation, cell transfusion, organ transplantation, organ preservation, cell preservation, asthma, irritable bowel disease, irritable bowel syndrome, ulcerative colitis, colitis, bowel disease, cancer, leukemia, ischemia-reperfusion injury, stroke, neointimal thickening associated with vascular injury, bullous pemphigoid, neonatal obstructive nephropathy, familial hypercholesterolemia, atherosclerosis, dyslipidemia, aortic aneurisms, arteritis, vascular occlusion, including cerebral artery occlusion, complications of coronary by-pass surgery, myocarditis, including chronic autoimmune myocarditis and viral myocarditis, heart failure, including chronic heart failure (CHF), cachexia of heart failure, myocardial infarction, stenosis, restenosis after heart surgery, silent myocardial ischemia, post-implantation complications of left ventricular assist devices, thrombophlebitis, vasculitis, including Kawasaki's vasculitis, giant cell arteritis, Wegener's granulomatosis, traumatic head injury, post-ischemic-reperfusion injury, post-ischemic cerebral inflammation, ischemia-reperfusion injury following myocardial infarction and cardiovascular disease.   
     
     
         16 . The method according to any of  claims 12 - 15 , wherein the level of inflammation is decreased by at least 20% compared to the level of inflammation in the patient before being administered the pharmaceutical composition. 
     
     
         17 . The method according to  claim 16 , wherein the level of inflammation is measured by cellular infiltration, cytokine levels, pain scores, degree of swelling, pulmonary function, degree of bronchorelaxation, occurrence or level of abdominal complaints, or other chemical or clinical assessments. 
     
     
         18 . A kit comprising a unit dose of a compound according to any one of  claims 1 - 9 , in an appropriate container.